Paper in Stem Cell Reports

“Efficient designer nuclease-based homologous recombination enables direct PCR screening for footprintless targeted human pluripotent stem cells”, Stem Cell Reports 2014

Genetic engineering of human induced pluripotent stem cells (hiPSCs) via customized designer nucleases has been shown to be significantly more efficient than conventional gene targeting, but still typically depends on the introduction of additional genetic selection. This genome engineering has now been further optimized by scientists from the LEBAO, together with scientists from the departments of Experimental Hematology and Human Genetics (both MHH), as well as with cooperation partners from the University Medical Center Freiburg, as published in “Stem Cell Reports” in January 2014. In this study, the authors demonstrate a selection-independent gene targeting without the typically required antibiotic selection. This optimized approach allows targeted transgene integration into safe harbor sites for more predictable and robust expression and enables the introduction of mutations on nucleotide level as well as the gene correction of disease-specific mutations in patient-derived iPSCs. This method is not only of utmost importance for the investigation of disease mechanisms and for the generation of disease-corrected, patient-derived iPSC lines for research purposes but, ultimately, for future clinical cell therapeutic applications of genetic diseases like cystic fibrosis.

Merkert, S., Wunderlich, S., Bednarski, C., Beier, J., Haase, A., Dreyer, A. K., Schwanke, K., Meyer, J., Gohring, G., Cathomen, T., and Martin, U. 2014. Efficient designer nuclease-based homologous recombination enables direct PCR screening for footprintless targeted human pluripotent stem cells. Stem Cell Reports 2, no. 1:107